The Functions of BRCA2 in Homologous Recombinational Repair
Abstract
The BRCA2 breast cancer gene is a major contributor to a dominantly inherited predisposition to the disease. Recent evidence has indicated that the critical functions of BRCA2 associate with chromosome stability and with DNA repair medicated by homologous recombination. Using homologous recombination assay, we have demonstrated that disruption of the BRCA2- Rad51 interaction inhibits the HRR frequency in vivo. The results indicate that the interactions between Rad5l and BRCA2 are important for regulating HRR. The 6xHis-tagged baculoviral construct of BRC1-4, BRC5-8 and BRC1-8 fragments of bRCA2 have been made and individually expressed in insect cells. Each protein was purified using a Ni-NTA column and the identity of each protein was confirmed by Western analysis. We have used three assay systems to determine the activity of Rad5lB and Rad5lC proteins, including DNA binding, ATPase and DNA strand transfer.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jul 01, 2003
- Accession Number
- ADA420343
Entities
People
- David J. Chen
Organizations
- University of California, Berkeley